The Antioxidant Capacity Of Trans-anethole And The Potential Mechanism In Animals

Star anise has long been used as medicinal and edible plant in China.Star anise oil(SAO)was essential oil extracted from star anise by hydro distillation.Trans-anethole was the main active compound in star anise and SAO.Previous studies have found that trans-anethole had an excellent antioxidant capacity in preventing lipid oxidation in feed,improving the antioxidant capacity in birds and mammals and so on.However,there is little information about the potential mechanism of antioxidant capacity of SAO in animals.The key producer of reactive oxygen species(ROS)in many cells was nicotinamide adenine dinucleotide phosphate(NADPH)oxidase,which actively communicate during the host response to a wide variety of stimuli,including bacterial infections and foreign particles.The ROS not only help in the clearance of pathogens and foreign particles but also lead to inflammation and tissue oxidative injury.Three experiments were conducted to investigate the antioxidant capacity of trans-anethole and the potential mechanism in animals.1)Mice were used as experimental animal to investigate the antioxidant capacity of trans-anethole and the potential mechanism in mammals.2)SPF chickens were used as experimental animal to investigate the antioxidant capacity of trans-anethole and the potential mechanism in birds.3)The oxidative stability as well as application in broilers of trans-anethole were studied to investigate the practical application in animal production1 The antioxidant capacity of trans-anethole and the potential mechanism in miceEffects of trans-anethole on antioxidant capacity of mice:A total of 100 mice were randomly divided into 5 groups with 20 mice each after adaptive feeding for 7 days.The tested groups were supplemented with 100,200,400 and 800 mg/kg SAO respectively on the basis of the control.Supplementation of SAO significantly increased(P<0.05)the activities of glutathione peroxidase(GSH-Px)and catalase(CAT)in serum and jejunum,as well as CAT in liver,but reduced(P<0.05)the malonaldehyde(MDA)and protein carbonyl contents as compared to those in controlTrans-anethole attenuates lipopolysaccharides(LPS)induced oxidative injury in mice:A total of 120 mice were randomly divided into 6 groups with 20 mice each after adaptive feeding for 7 days.T1,Control;T2,LPS;T3,LPS+100 mg/kg SAO;T4,LPS+200 mg/kg SAO;T5,LPS+400 mg/kg SAO;T6,LPS+200 mg/kg ascorbic acid(VC).1)LPS induced a marked oxidative injury evident by decrease in activity of CAT and rise in contents of MDA and protein carbonyl in serum,liver and jejumun of mice(P<0.05).Levels of nuclear transcription factor-?B(NF-?B),interleukin-1?(IL-1?),interleukin-6(IL-6),interleukin-8(IL-8),and tumor necrosis factor-?(TNF-?)in serum were significantly increased(P<0.05)in LPS challenged mice.2)Supplementation of SAO significantly increased(P=0.040)body weight(BW)compared with those in the LPS group.Meanwhile,the mice in SAO groups had lower(P<0.001)levels of NF-?B,IL-1?,IL-8 and TNF-?Supplementation of SAO significantly increased(P<0.05)the activities of antioxidant enzymes but reduced(P<0.05)the MDA and protein carbonyl contents in serum,liver and jejunum of mice as compared with those in the LPS group,and the efficacy of SAO in mice was dose dependent.3)Compared to the LPS group,the mRNA and protein expression levels of NF-erythroid-2-related factor 2(Nrf2)in liver and jejunum were significantly increased(P<0.05),as well as the mRNA expression levels of SOD1,CAT and GPX4 in SAO supplemented groups.These findings suggested that the mechanism by which SAO exerted its antioxidant capacity in mice was correlated with the activation of Nrf2-ARE signaling pathway.SAO attenuated LPS induced oxidative injury possibly by preventing cytotoxic effects of oxidative products and inflammation cytokines2 The antioxidant capacity of trans-anethole and the potential mechanism in SPF chickensEffects of trans-anethole on antioxidant capacity of SPF chickens:A total of 384 thirty-one-week-old SPF chickens were randomly divided into 4 treatments with 8 replicates respectively.Chickens were fed with the basal diet supplemented with 0(control),200,400 and 600 mg/kg SAO,respectively.1)The BW and average daily feed intake(ADFI)in birds were similar(P>0.05)among all the groups.2)Supplementation of SAO significantly increased(P<0.05)the activities of GSH-Px and CAT but reduced(P<0.05)the MDA and protein carbonyl contents in serum,liver and jejunum of birds as compared with those in the control,and the efficacy of SAO in birds was dose dependent.3)Compared to the control group,birds fed diets supplemented with SAO significantly increased(P<0.05)the mRNA and protein expression levels of Nrf2 in liver and jejunum,as well as the mRNA expression levels of CAT and GPX4.The enhancement of antioxidant capacity by SAO in birds might be through upregulation of Nrf2-ARE signaling pathwayTrans-anethole attenuates Escherichia coli(E.coli)induced inflammation and oxidative injury in SPF chickens:A total of 480 thirty-one-week-old SPF chickens were randomly divided into 5 treatments with 8 replicates,respectively.T1,Control;T2,E.coli;T3,E.coli+200 mg/kg SAO;T4,E.coli+400 mg/kg SAO;T5,E.coli+600 mg/kg SAO.1)Compared to the control group,BW was significantly decreased(P=0.016)in the E.coli group.E.coli challenge increased(P<0.001)the relative mRNA expression levels of NF-?B.IL-1?,IL-6,IL-8 and TNF-? in liver and jejunum in comparison with the control group Meanwhile,E.coli challenge decreased(P<0.05)activity of GSH-Px but increased(P<0.05)contents of MDA and protein carbonyl in serum,liver and jejunum of birds.Those results showed that E.coli challenge induced inflammation and oxidative injury in birds.2)Supplementation of SAO significantly decreased(P<0.001)the mRNA expression levels of inflammatory related genes in liver and jejunum when compared to the E.coli group Supplementation of SAO could also increase(P<0.05)activities of GSH-Px and CAT but decrease(P<0.05)content of protein carbonyl in serum,liver and jejunum of birds when compared to the E.coli group.4)Compared to the E.coli group,the mRNA and protein expression levels of Nrf2 in liver and jejunum were significantly increased(P<0.05)in SAO supplemented groups,as well as the mRNA expression levels of SOD2?GPX4 and CAT Supplementation of SAO could attenuate E.coli challenge induced inflammation and oxidative injury,the mechanism by which SAO exerted its antioxidant capacity in birds was correlate with the activation of Nrf2-ARE signaling pathwayEffects of trans-anethole on gut microbiome of SPF chickens during E.coli challenge:The experimental design was the same as above.1)E.coli challenge significantly increased(P<0.05)the contents of E.coli in duodenum,jejunum,ileum and cecum,but reduced(P<0.05)the relative abundances of Parabacteroides,unidentified Rurninococcaceae and Rikenella in cecum in comparison with the control group.2)Compared to the E.coli group,the contents of E.coli in intestines were significantly decreased(P<0.05)but the relative abundance of Parabacteroides in cecum was significantly increased(P<0.05)in SAO supplemented groups3 The oxidative stability of trans-anethole as well as its application in broilersThe oxidative stability of trans-anethole:The antioxidant capacity of SAO was evaluated based on its scavenging abilities of 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)(ABTS)and 1,1-diphenyl-2-picryIhydrazyl(DPPH)free radicals.Ascorbic acid(VC)was used as a reference standard.The results showed that the scavenging ability of ABTS free radical in SAO was significantly higher(P<0.001)than the VC control.Experiment was then conducted to investigate the effects of storage temperature and sealed condition on oxidative stability of SAO in vitro.Storage temperatures at 4? of 20? and sealed conditions at sealed and unsealed were used in a 2 x 2 factorial arrangement in randomized complete blocks.Results showed that storage temperatures significantly changed(P<0.001)scavenging ability of ABTS free radical in SAO during 4 wk of storage.Meanwhile,sealed conditions significantly changed(P<0.05)scavenging abilities of ABTS and DPPH free radicals of SAO during 2 and 6 wk of storage.The antioxidant capacity of SAO in vitro can be influenced by the storage conditions(temperatures,sealed conditions and time),which was more suitable for storage at 4?,sealed condition.Application in broilers:A total of 288 1-day-old broilers were randomly assigned to 3 treatments with 8 replicates of 12 birds each.The treatments were corn-soybean meal basal diet supplemented with 5000 mg/kg star anise and 220 mg/kg SAO of diet.The composition of trans-anethole in star anise and SAO supplemented groups was equal,to meet a level of 207±2.00 mg/kg.1)Birds supplemented with star anise and SAO had greater(p<0.05)BW at 21 d and 42 d of age than control birds.Broilers supplemented with SAO had higher(P<0.05)ADFI during the grower period(22-42 d),as well as ADFI and average daily gain(ADG)during the entire period(1-42 d)than those in the control group.2)Supplementation of star anise and SAO significantly increased(P<0.05)activity of GSH-Px in serum and liver of broilers at 21 d and 42 d of age,as compared with those in the control group.Meanwhile,supplementation of SAO also increased(P<0.05)activities of total superoxide dismutase(T-SOD)and CAT but decreased(P<0.05)MDA content in serum and liver of broilers at 42 d of age,as compared with those in the control group.3)Among all the groups,muscles of SAO supplemented group appeared to contain the lowest(P=0.012)boiling loss but highest(P<0.001)concentration of inosinic acid.Dietary supplementation of star anise and SAO(the composition of trans-anethole was equal)improved growth performance and antioxidant abilities.Meanwhile,the effect of SAO in broilers was better than star anise.In conclusion,these results above indicated that trans-anethole exhibited a strong antioxidant capacity in animals.The mechanism by which trans-anethole exerted its antioxidant capacity in mice was correlated with the activation of Nrf2-ARE signaling pathway.Trans-anethole attenuated LPS induced oxidative injury in mice possibly by preventing cytotoxic effects of oxidative products.The mechanism by which trans-anethole exerted its antioxidant capacity in birds was also correlated with the activation of Nrf2-ARE signaling pathway.Meanwhile,supplementation of trans-anethole could attenuate E.coli challenge induced inflammation and oxidative injury by preventing cytotoxic effects of oxidative products and inflammation cytokines.